for

Min-Ho Lee mhlee at WWW.BRIC.POSTECH.AC.KR
Thu Dec 26 01:04:35 EST 1996


Shou Jiang wrote:
> 
> Hi, Netters,
> 
> Who would like to offer more effective methods to extract lambda DNA from
> lambda library? any kit exist?
> 
> Thanks in advance.
> 
> Please reply to shouj at nic.bmi.ac.cn
> 
> Shou Jiang
Hi!
We collected this subject in our web page
(http://bric.postech.ac.kr/http://bric.postech.ac.kr/forum/bric-forum/newsgroup/bio_methods/)
as follows;

Subject: Re: Lambda DNA extraction protocol
Date: Wed, 04 Dec 1996 | From: robin_beech at maclan.mcgill.ca (Robin
Beech)

slee at pbi.nrc.ca ("Lee, Sun") wrote: Would anyone like to introduce me a
really simple way for this? Most of published methods that were armed
with 'easy' , 'simple' .... were
actually not like that. Looking for in -hand method that really works. 

Lambda miniprep

     1) Grow an overnight liquid lysate
     2) Spin down 1ml of lysate to remove bacterial debris
     3) Transfer 0.8ml of lysate to a new eppendorf
     4) Add DNAse to 1ug per ml and incubate at RT for 15 min
     5) Add 0.2ml of Tris (50mM pH 8.0) EDTA (10mM) SDS (5%) shake
gently and incubate 70 deg for 15 min
     6) Cool on ice and add 0.15ml 8M potassium acetate
     7) Spin down the protein precipitate
     8) Phenol-chloroform extract the supernatent
     9) Add 2.5 vol of ethanol or 1 vol of isopropanol to precipitate
the DNA

This can also be scaled up to process 8ml of lysate Hope this helps -
good luck

Robin N. Beech
Institute of Parasitology
McGill University

------------
Min-Ho Lee ; mhlee at bric.postech.ac.kr
Web for the Korean Entomologist (my home page) : 
http://bric.postech.ac.kr/about/mhlee/
BRIC, POSTECH
http://bric.postech.ac.kr



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